Page 1/25 Ocimum Sanctum Ameliorates Valproic Acid – Induced Hepatotoxicity Muhammad Omer Iqbal ( [email protected]) Ocean University of China School of Medicine and Pharmacy https://orcid.org/0000-0002-7122-9422 Qurratulain Rahim The Islamia University of Bahawalpur Pakistan Department of Pharmacy Hania Mehboob Khan The Islamia University of Bahawalpur Pakistan Department of Pharmacy Imran Akhter The Islamia University of Bahawalpur Pakistan Department of Pharmacy Faiza Naeem Lahore College for Women University Faculty of Pharmaceutical and Allied Health Sciences Fiaz ud din Ahmad islamia university of bahawalpur Research Keywords: Ocimum sanctum, Flavonoids, Valproic acid induced hepatotoxicity, Lamiaceae, Antioxidant activity Posted Date: December 30th, 2020 DOI: https://doi.org/10.21203/rs.3.rs-136212/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License
AbstractOBJECTIVE: Evaluation of hepatoprotective potentials of Ocimum sanctum against Valproic-acid-induced-hepatotoxicity in Wistar-albino-rats.
METHOD: 70% of ethanolic extract of Ocimum sanctum was prepared under reduced pressure of rotaryevaporator. Wistar albino rats were used as the experimental model and rats were divided into fourgroups (six animals each). The normal group received normal saline and group 2, 3 and 4 was injectedvalproic acid (500mg/kg) for four consecutive days respectively. Group 1 and 2 received normal salinethroughout the period of study about 21 days while group 3 and 4 received different doses of extract ofOS i.e. 200mg/kg and 300mg/kg. Through retro-orbital blood samples were collected on alternative dayssuch as 0,7,21. By using one-way ANOVA, data was analyzed. Hepatotoxicity induced by valproic acid atthe dosage of (500mg/kg) resulted in signi�cant elevation in weight of animals and serum hepaticenzymes level of ALAT, ASAT, ALP and increase in the serum bilirubin.
RESULTS: OS at different doses (200mg/kg and 300mg/kg) considered statistically signi�cant (p ≤ 0.05)against all parameters. OS cause a signi�cant reduction in weight of animals and serum enzymesbiomarkers i.e. (ALAT, ASAT and ALP) including bilirubin content. OS may prove its hepatoprotectiveactivity by increase a signi�cant level of protein albumin.
CONCLUSION: antioxidant activity of OS and secondary metabolites such as �avonoids depictshepatoprotective nature against valproic-acid-induced-hepatotoxicity.
IntroductionIn year 1882, Valproic acid (2-propyl-pentanoic acid, 2 propylvaleric acid) was �rst synthesized by Burton[1]. In 1963, VPA (valproic acid) was used as a molecular carrier in anti-convulsive activities. Itspharmacological activities demonstrated that VPA was used in rodents against pentylenetetrazol-inducedconvulsions [2]. Today, worldwide valproic acid is used as an anti-epileptic drug to treat several forms ofepilepsy and various types of seizures, that affecting both children’s and adults [3]. The major undesirableside effects of VPA, included teratogenicity and hepatotoxicity [4]. Patients receiving valproic acid,causing hepatic failure resulting in fatalities [5]. Changes in serum cholesterol, triglycerides, fat glucoseand gain of weight, have also been associated with side effects of VPA [6].
In rat hepatocytes, VPA is associated with biochemical disturbances such as inhibition of fatty acidoxidation, ketogenesis, gluconeogenesis, urea synthesis and reduced level of acetyl-co A (Becker andHarris, 1983) [7]. Administrating high dose of VPA consistently, about 500-750mg/kg produces microvesicular steatosis. The steatogenic effect is produced because of toxic metabolites of VPA. Mechanismof these effects is due to (a) depress the synthesis of proteins; (b) through mitochondrial injurydepressing oxidation of fatty acids, or (c) enhancing mobilization to the liver from fat depots [8].Medicinal plants as an antioxidant property of �avonoids and a tendency to protect the membrane, is
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observed as a hepatoprotective medicinal plants [9] Some hepatoprotective medicinal plants includes, Azadirachta indica, Cassia �stula, Silybum marianum [10].
Ocimum sanctum commonly known as “Tulsi”. The word tulsi is a Sanskrit word which means “theinimitable one” [11]. In bygone literature, tulsi describe as “queen of herbs” [12]. In Ayurvedic and Unanisystem of medicines ocimum sanctum is also known as “Sweet Basil or Holy Basil” [13]. Worldwide,about 150 species or class of Ocimum is present but basil is the real one who developed fundamentally[14]. Plant of tulsi is about 30-60 cm tall with hairy stem. Leaves are green having petioles and up to 5 cmlong. Ocimum sanctum is a �avoring remedy and enhanced many basic ailments [15].Flavonoids,Alkaloids, Saponins, Tanins, Phenols, Anthocynins, Terpenoids and Steroils [16]. The hydro-ethanolic extract of OS leaves, orally administered at 200mg/kg dose or more than 200 mg/kg againstccl4, acetaminophen and lead-induced hepatotoxicity in albino rats, shows that OS induced protectiveeffect in these animal models. Ocimum sanctum reduced the level of serum hepatic enzymes, such asALAT, ASAT and ALAP in above animal models [17]. OS also induced hepatoprotective effect against anti-tubular drugs-induced hepatotoxicity in albino rats [18].
PLANT ANATOMY
Kingdom : Plantae
Division : Magnoliophyta
Class : Magnoliopsida
Order : Lamiales
Family : Lamiaceae
Genus : Ocimum
Species : O.tenui�orum
Botanical Name : Ocimum Tenui�orum (Singh et al., 2012a). [9].
IPNI Life Sciences Identi�er (LSID)
urn:lsid:ipni.org:names:30252269-2
Kew Names and Taxonomic BackboneThe International Plant Names Index and World Checklist of Selected Plant Families.
Research MethadologyEthical consideration
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The study was approved by IRB of The Islamia University of Bahawalpur, Bahawalpur, Pakistan. Date ofapproval of this study was 03 September 2016 under the approval number of IUB/03-16/34059.
Material and Equipment’s
For research purpose chemicals of analytical grade were used included valproic acid (platinumpharmaceutical LTD), ketamine (Indus pharma Lahore), xylazine (prix pharmaceutical Lahore), ether,aqueous ethanol all one of analytical grade.
Wistar Albino rats weighting 220-270g were taken and kept in cages. Cages with polycarbonate layersecured by raw dust, under standard lab conditions it changed after every three days (temperature:27±2 c in animal house of pharmacology research lab, faculty of pharmacy and alternative medicine, theIslamia university of Bahawalpur. Rats were provided with standard diet pallets and water ad libitum.Wistar Albino rats were acclimatized to a regimen of 12 hours of light and 12 hours of haziness. Byethical committee of the Islamia University of Bahawalpur, study was approved. According to studyanimal were divided.
Subject of study:
24 Wistar rats of either sex were assigned into four groups each having six animals and categorized as;
Group 1 was named as normal control and given normal saline.
Group 2 was named as valproic acid group and given (500 mg /kg)
Group 3 was named as VPA + extract 1 and given (200 mg/kg) of O.S extract.
Group 4 was named as VPA + extract 2 and given (300 mg/kg) of O.S extract
Plant collection:
Fresh, Ocimum Sanctum leaves were purchased from local nursery of Bahawalpur, Pakistan on May2017. Information was collected about the medicinal uses of plants to treat hepatic injury.
Preparation of Crude Extract:
The leaves of Ocimum Sanctum Linn were separated, dried under shade for 15 days and powdered in ablender. It was grinded by electric grinder and then soaked in a mixture of 70% v/v ethanol and 30% v/vdistilled water for three days with shaking and agitation occasionally. By muslin fabric, soaked materialwas separated for coarse �ltration and afterward by Whattman �lter paper (NO.1).
Then by using rotary evaporator (Heidolph Laborota 4000 e�cient, Germany) at a temperature of 30-40 C the �ltrate obtained was then evaporated under reducedpressure. Until the�ltrate become semi solid the process of evaporation was Continue and placed the �ltrate into theincubator for further drying then stored in a refrigerator (-4 c) till further use.
Induction of Hepatotoxicity:
Valproic acid (500 mg/kg orally) once a day for four consecutive days was used to induce hepatotoxicity.
Experimental Protocol
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Rats were assigned into four groups randomly and each group having six animals. Group-1 was normalcontrol and animals were given normal saline as a vehicle for 21days. Group-2 groupwas intoxicated and toxicity was caused by valproic acid (500 mg/kg) once daily for four consecutivedays given orally and received normal saline after further for 21 days. Group-3 was considered treatmentgroup and treated with Ocimum sanctum (200 mg/kg) dose orally for 21 days and received (500 mg/kg)orally valproic acid for four consecutive days. Group-4 was considered treatment group and treated withOcimum sanctum at dose (300 mg/kg) orally for 21 days and received valproic acid (500 mg/kg) orallyfor four consecutive days. The blood was collected via retro-orbital at day 0, 7 and 21 and serum wasseparated by centrifugation at 60 rpm about 20 minutes. Animals were sacri�ced at the end of theexperimental period.
Biochemical Parameters Studied
All parameters were carried out by human diagnostic kit Germany.
Liver Enzyme Test
Liver Protein Test
Bilirubin
Statistical Analysis
The obtained data were expressed as mean ±S.E.M. The data were subjected to one-way (ANOVA)Bonferroni’s post hoc test for statistical signi�cance between experimental groups. P-values <0.05 wereexamined as statistically signi�cant.
ResultsResults of control, VPA and VPA+O.S (200mg/kg) treated group and VPA+ O.S (300mg/kg) treated group
Effect of O.S on Body weight
Variance in the body Weight of rats (all groups) was observed on day 0, 7 and 21 during the period ofthree weeks study. It was observed that the body weight among all groups at day “0” was not signi�cantlydifferent (all p>0.05). As the study continue, it was observed that on day “7” and “21” body weight ofcontrol group was signi�cant higher as compared to the body weight of control group at day “0” (allp<0.05). Generally, incredibly the body weight of VPA treated group was showed signi�cant increase onday “7” and day “21” as compared to the body weight at day 0(all p<0.05). With the progression of days,VPA+O.S(200mg/kg) treated group and VPA+O.S(300mg/kg) treated groups of animals, exhibitedsigni�cant reduction in body weight on day “7” and day “21” as compared to the body weight of VPAtreated group on day 7 and day 21 respectively(allp<0.05).
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Table 1: Effects of crude extract of O.S on body weight in rats treated with VPA
Liver damage assessment was done by biochemical investigation of serum Alkaline phosphate (serumALP) after three weeks of study. The ALP level was calculated on day “0”, “7” and “21” respectively. Theobservations made no signi�cant difference at day “0” among all groups (all p>0.05). It was observedthat on day “7” and day “21” the level of control group was not signi�cantly different as compared to thecontrol at day 0” (all p>0.05). As the study proceeded, it was analyzed that the ALP level of VPA treatedgroup on day “7” and “21” was signi�cant increased as compared to the VPA group at day 0(all p<0.05).Interestingly, signi�cant reduction in ALP level of VPA+O.S (200mg/kg) and VPA+ O.S (300mg/kg treatedgroups on day 7 and 21 compared to the VPA treated group at day 7 and 21 respectively (all p<0.05).Prevention on the decrease in ALP levels at the dose 200 and 300 mg/kg was found highly signi�cant.
Table 2: Effects of crude extract of O.S on Alkaline phosphate (ALP) in rats treated with VPA
DAYS OF OBSERVATIONS
Parameters Groups 0 DAY 7th DAY 21 DAY
ALP
(u/l)
Control 89 ± 6.5 107 ± 4.2 120 ± 6.8
VPA (500mg/kg) 97 ± 9.1 225 ± 9.7(*) 226 ± 5.1(*)
VPA+O.S
(200mg/kg)
77 ± 3.7 194 ± 5.3(&) 172 ± 5.3(&)
VPA+O.S
(300mg/kg)
80 ± 3.4 179 ± 5.9(#) 160 ± 6.8(#)
Effect of O.S on Serum Alanine Aminotransferase (ALAT)
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Liver damage evaluation was done by biochemical examination of serum Alanine Aminotransferase(serum ALAT) following three weeks of study. The level of ALAT was calculated on day “0”, “7” and “21”respectively. No signi�cant difference at day “0” among all groups when observations were made (allp>0.05). It was noticed that on day “7” and day “21” the level of control group was not signi�cantlydifferent as compared to the control at day 0” (all p>0.05). As the study proceeded, it was evaluated thatthe ALAT level of VPA treated group on day “7” and “21” was signi�cant increased as compared to theVPA group at day 0(all p<0.05). Incredibly, signi�cant reduction in ALAT level of VPA+O.S (200mg/kg) andVPA+ O.S (300mg/kg treated groups on day 7 and 21 compared to the VPA treated group at day 7 and 21respectively (all p<0.05). Anticipation on the decrease in ALAT levels at the dose 200 and 300 mg/kg wasbegun highly signi�cant.
Table 3: Effects of crude extract of O.S on Alanine Aminotransferase (ALAT) in rats treated with VPA
DAYS OF OBSERVATIONS
Parameters Groups 0 DAY 7th DAY 21 DAY
ALAT
(u/l)
Control 29 ± 3.5 33 ± 4.5 30 ± 2.5
VPA (500mg/kg) 28 ± 3.5 98 ± 4.9(*) 97 ± 8.9(*)
VPA+O.S
(200mg/kg)
23 ± 3.6 74 ± 6.3(&) 70 ± 4.2(&)
VPA+O.S
(300mg/kg)
30 ± 3.0 60 ± 6.9(#) 58 ± 6.0(#)
Effect of O.S on Serum Aspartate Aminotransferase (ASAT)
Liver anguish assessment was done by biochemical estimation of serum Aspartate Aminotransferase(serum ASAT) after three weeks of study. The ASAT level was calculated on day “0”, “7” and “21”respectively. The observations made no signi�cant difference at day “0” among all groups (all p>0.05). Itwas analyzed that on day “7” and day “21” the level of control group was not signi�cantly different ascompared to the control at day 0” (all p>0.05). As the study proceeded, it was noticed that the ASAT levelof VPA treated group on day “7” and “21” was signi�cant increased as compared to the VPA group at day0(all p<0.05). Interestingly, signi�cant reduction in ASAT level of VPA+O.S (200mg/kg) and VPA+ O.S(300mg/kg treated groups on day 7 and 21 compared to the VPA treated group at day 7 and 21respectively (all p<0.05). Deterrence on the decrease in ASAT levels at the dose 200 and 300 mg/kg wasestablished highly signi�cant.
Table 4: Effects of crude extract of O.S on Aspartate Aminotransferase (ASAT) in rats treated with VPA
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DAYS OF OBSERVATIONS
Parameters Groups 0 DAY 7th DAY 21 DAY
ASAT
(u/l)
Control 79 ± 4.3 73 ± 6.4 76 ± 3.8
VPA (500mg/kg) 63 ± 5.5 180 ± 11(*) 171 ± 6.4(*)
VPA+O.S
(200mg/kg)
71 ± 7.8 137 ± 11(&) 140 ± 5.6(&)
VPA+O.S
(300mg/kg)
69 ± 8.4 25 ± 12(#) 116 ± 6.0(#)
Effect of O.S on Serum Albumin
Liver injury assessment was done by biochemical evaluation of serum Albumin following three weeks ofstudy. Level of Albumin was calculated on day “0”, “7” and “21” respectively. The observations made nosigni�cant difference at day “0” among all groups (all p>0.05). It was observed that on day “7” and day“21” the level of control group was not signi�cantly different as compared to the control at day 0” (allp>0.05). As the study proceeded, it was analyzed that the Albumin level of VPA treated group on day “7”and “21” was signi�cant decreased as compared to the VPA group at day 0(all p<0.05). Generally,signi�cant increase in Albumin level of VPA+O.S(200mg/kg) and VPA+ O.S(300mg/kg treated groups onday 7 and 21 compared to the VPA treated group at day 7 and 21 respectively (all p<0.05). Precaution onthe increase in Albumin levels at the dose 200 and 300 mg/kg was created highly signi�cant.
Table 5: Effects of crude extract of O.S on Albumin in rats treated with VPA
Liver damage assessment was done by biochemical examination of serum total bilirubin (serum TBR)after three weeks of study. The level of TBR was calculated on day “0”, “7” and “21” respectively. The
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observations made no signi�cant difference at day “0” among all groups (all p>0.05). It was observedthat on day “7” and day “21” the level of control group was not signi�cantly different as compared to thecontrol at day 0” (all p>0.05). As the study continue, it was analyzed that the TBR level of VPA treatedgroup on day “7” and “21” was signi�cant increased as compared to the VPA group at day 0(all p<0.05).Generally, incredibly signi�cant reduction in TBR level of VPA+O.S(200mg/kg) and VPA+ O.S(300mg/kg)treated groups on day 7 and 21 compared to the VPA treated group at day 7 and 21 respectively (allp<0.05). Prevention on the decrease in TBR levels at the dose 200 and 300 mg/kg was begun highlysigni�cant.
Table 6: Effects of crude extract of O.S on TBR in rats treated with VPA
Liver damage assessment was done by biochemical examination of serum direct bilirubin (serum DBR)after three weeks of study. The level of DBR was calculated on day “0”, “7” and “21” respectively. Theobservations made no signi�cant difference at day “0” among all groups (all p>0.05). It was observedthat on day “7” and day “21” the level of control group was not signi�cantly different as compared to thecontrol at day 0” (all p>0.05). As the study continue, it was analyzed that the DBR level of VPA treatedgroup on day “7” and “21” was signi�cant increased as compared to the VPA group at day 0(all p<0.05).Generally, incredibly signi�cant reduction in DBR level of VPA+O.S(200mg/kg) and VPA+ O.S(300mg/kg)treated groups on day 7 and 21 compared to the VPA treated group at day 7 and 21 respectively (allp<0.05). Prevention on the decrease in DBR levels at the dose 200 and 300 mg/kg was begun highlysigni�cant.
Table 7: Effects of crude extract of O.S on DBR in rats treated with VPA
DiscussionThe objective of wild medicinal or therapeutic plants to treat human diseases is not recent. Since ancienttime herbal medicine played an important role in curing diseases of humans [19]. Many environmentalchemicals and drugs can cause some degree of injuries and leading cause of acute liver injury, is a drug-induced liver injury [20]. Generally, drugs can cause an imbalance between prooxidant and antioxidantlevels in cells and tissues. Reactive oxygen species (ROS) are prooxidants which can cause tissue injury[21]. several medicinal plants with their antioxidant properties are effective to cure injuries of liver andbene�cial as a hepatoprotective plant. Present study was designed to investigate the hepatoprotectiveeffect of ocimum sanctum linn against valproic acid-induced hepatotoxicity in albino rats. Valproic acid(2-propyl-pentanoic acid, 2 propyl valeric acid), used as an anti-epileptic drug to treat several forms ofepilepsy and various types of seizures [3]. The major undesirable side effects of VPA, includedteratogenicity and hepatotoxicity [4]. In VPA-induced liver injury, pathogenesis involves the toxicmetabolites of drug that effect directly the cell biochemistry or evoke an immune response. β-oxidationan important pathway for oxidative metabolism. In rat hepatocytes, VPA is associated with biochemicaldisturbances such as inhibition of fatty acid oxidation, ketogenesis, gluconeogenesis, urea synthesis andreduced level of acetyl-co A [22]. Valproyl-co A, helps to form esters with fatty acids [23]. A reactivemetabolite of valproic acid that is 2,4-Diene-valproyl-CoA cause inhibition and inactivation of β-oxidationenzymes [24]. Several studies investigated that treatment with VPA, in case of liver injury, is associatedwith oxidative stress in many patients and in animal models too [25]. The etiology of oxidative stress ishypothesized because of overproduction of reactive oxygen species (ROS) and/or reduced antioxidantcapacity [26]. Glutathione (GSH) is an important antioxidant cellular biomolecule. A reactive metabolite ofVPA that is 4-ene-VPA cause depletion of GSH in rat hepatocytes and induced oxidative stress [27].Valproic acid-induced hepatotoxicity showed by decrease or increase in the level of serum hepaticproteins and enzymes test [28]. So, my study was designed to examine the hepatotoxic effects of valproicacid in albino rats.
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Ocimum sanctum play an important role in various pharmacological actions. Traditionally ocimumsanctum is ful�l actions in many other prevention and cure of illness like fever and common cold,coughs, sore throat, kidney stone, children’s Ailments, mouth infection, insect bites, skin disorders, teethdisorders, headaches and colic pain [29]. OS shows antidote activity against several poisons [30]. OSpotentiate signi�cant effect to lower of sugar level in blood. It exerts hypoglycemic effect instreptozotocin-induced diabetic rats [31]. Moreover, ocimum sanctum induced their hepatoprotectiveactivity in various animal models against CCl4, acetaminophen, lead and anti-tubular drugs [32]. Antioxidant property of ocimum sanctum leaves have studied already. Chemical toxins and radiationinduce biological damage because of forming reactive oxygen species, such as singlet oxygen andsuperoxide’s, hydroxyl and hydroperoxyl radicals, hydrogen peroxide and organic peroxides. Extract of OSleaves and their �avonoids, such as orientin and vicenin have shown high antioxidant activity in vivo andanti-lipid peroxidative effect in vitro [33].
However, the antioxidant potential of herbs is due to the redox properties of their phenolic compounds,which allow them to act as a reducing agent, hydrogen donors and singlet oxygen species [34]. Phenoliccompounds are secondary metabolites that possess high antioxidant activity [35]. It is stronglysuggesting that free radicals scavenging a major mechanism, through which ocimum sanctum protectagainst tumor induction and cellular damage [36]. Also, ocimim sanctum reduced erythrocyte lipidperoxidation activity induced against hypercholesterolemia in albino rats and protect aortic and livertissues against hypercholesterolemia-induced peroxidative damage [37].
Previous study has suggested that the plant extract contains constituents having hepatoprotective andstrong antioxidant secondary metabolites that counteract the lethal impacts of valproic acid. So, mystudy was designed to determine hepatoprotective action of ocimum sanctum extract against valproicacid-induced hepatotoxicity. Hepatotoxicity was observed through some invitro study and by various liverfunction parameters in-vivo. In invitro study, gain of weight was observed in VPA treated group animalsand weight was signi�cantly reduced when animals treated with ocimum sanctum extract. One sideeffect of VPA is weight gain and during VPA treatment the amount of average weight gain isapproximately 6 kg in human [38]. Invitro studies suggest that weight is gain when VPA initiatespancreatic insulin secretions that might increase energy storage and appetite. It concluded that possiblyinsulin resistance and hyperinsulinemia cause to weight gain [39]. Previous studies showed that inexperimental animals, ocimum sanctum leaves induce hypoglycemic effects. A study was conductedwith OS and neem stated that in diabetic patients this combination is good in lowering sugar level [40].OS potentiate signi�cant effect to lower of sugar level in blood. It exerts hypoglycemic effect instreptozotocin-induced diabetic rats.
The level of serum alkaline phosphatase (ALP), alanine aminotransferase (ALAT) and aspartateaminotransferase (ASAT) was increased after chronic administration of valproic acid in animals oftoxicities group. The level of above serum hepatic enzymes was signi�cantly reduced when extract ofocimum sanctum was given to treated group of animals. Previous studies showed that Valproic acidinduced liver injury and increase levels of serum hepatic enzymes [41]. In the cell cytoplasm ASAT, ALAT
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and ALP are located and after administration of ocimum sanctum extract improved toxicity and hasintense effect in restoration of AST, ALT and ALP and decreased level of these enzymes towards theirrespective normal values. The hepatic e�ciency improved, and regeneration of the hepatic tissue wasindicate by the restoration of the bilirubin levels in previous studies [42].
The most generous circulating protein in the plasma is albumin and the vital protein synthesized by theliver [43]. In my study valproic acid induced toxicity by decreasing the level of albumin in group ofanimals treated by valproic acid. The extract OS was given in treated group animals and study showsthat the level of albumin increased signi�cantly. In previous studies it was discussed that valproic acidinduced toxicity by decreasing the level of albumin [28]. Noteworthy, the level of albumin afteradministration of OS extract showed signi�cant increase in their levels in previous studies.
My study was exhibits that after chronic administration of valproic acid the level of serum bilirubin (totaland direct) was increased and shows that toxicity is induced in animals. In contrast the level wasdecreases and comes near to their normal values after giving ocimum sanctum extract in treated groupof animals. In previous studies, after induction of toxicity by valproic acid the level of serum bilirubin(total and direct) was increased signi�cantly compare to normal [28].
However, in vitro and in-vivo both studies show that valproic acid induced toxicities in hepatic enzymesand proteins biomarkers and disturbed their levels and in contrast of valproic acid, extract of ocimumsanctum induced its signi�cant effects to normalize the level of serum hepatic enzymes. However,advance research is needed to determine the exact mechanism of action. Also, whether hepatotoxicity ofvalproic acid is cumulative remains contentious.
ConclusionPresent study may have concluded that valproic acid induces hepatic damage in rat liver and causeoxidative stress in hepatocytes of rat. Reactive oxygen species altered the hepatic antioxidant enzymesand proteins and depletion of free radicals may cause injury of liver in albino rats. In contrast, ocimumsanctum linn due to its strong antioxidant activity protect the liver from damage. ocimum sanctum canminimize the effect of hepatotoxic drugs and other chemicals. Therapeutic potential of ocimum sanctumas a hepatoprotective medicinal plant has been proven. We endorse further studies and more clinicaltrials conducted to underprop its hepatoprotective potential and other therapeutic uses.
AbbreviationsOS: Ocimum Sanctum
B.W: Body weight
NaOH: Sodium hydroxide
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TCA: Trichloroacetic acid
VPA: Valproic acid
i.p: Intra peritoneal
ROS: Reactive oxygen species
BTW: Between
DeclarationsEthics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
Acknowledgements
Not applicable
Funding
Not applicable
Availability of data and materials
Not applicable
Authors’ Contributions
Omer Iqbal, Qurratulain Rahim, Fiaz ud din Ahmad designed the study. Qurratulain Rahim performed theexperiments and compiled the data. Faiza Naeem and Omer Iqbal analyzed the data and wrote themanuscript. Hania Mehboob Khan, Imran Akhtar, Quratulain Rahim, Omer Iqbal coordinated the projectpreparation of the �nal version of the manuscript.
Acknowledgment
The study was conducted by the Department of Pharmacy, the Islamia University of Bahawalpur.
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Funding: None
Con�ict of interest: None
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Figures
Figure 1
Tulsi
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Figure 2
Steps involved in preparation of crude extract of Ocimum Sanctum linn
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Figure 3
Body weight of normal control, VPA, VPA+O.S (200mg/kg), VPA+ O.S (300mg/kg). The values are mean ±SEM (n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferronipost hoc test for all gatherings in particular days. The results are considered signi�cant (p<0.05). *indicates p<0.05 vs normal. & indicates p<0.05 vs VPA.
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Figure 4
ALP of normal control, VPA, VPA+O.S (200mg/kg), VPA+ O.S (300mg/kg). The values are mean ± SEM(n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferroni posthoc test for all gatherings in particular days. The results are considered signi�cant (p<0.05). * indicatesp<0.05 vs normal. & indicates p<0.05 vs VPA. # indicates p<0.05 vs VPA.
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Figure 5
ALAT of normal control, VPA, VPA+O.S (200mg/kg), VPA+O.S (300mg/kg). The values are mean ± SEM(n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferroni posthoc test for all gatherings in particular days. The results are considered signi�cant (P<0.05). *indicatesp<0.05 vs normal. &indicates p<0.05 vs VPA. #indicates p<0.05 vs VPA.
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Figure 6
ASAT of normal control, VPA, VPA+O.S (200mg/kg), VPA+O.S (300mg/kg). The values are mean ± SEM(n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferroni posthoc test for all gatherings in particular days. The results are considered signi�cant (P<0.05). *indicatesp<0.05 vs normal. &indicates p<0.05 vs VPA. #indicates p<0.05 vs VPA.
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Figure 7
Albumin of normal control, VPA, VPA+O.S (200mg/kg), VPA + (300mg/kg). The values are mean ± SEM(n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferroni posthoc test for all gatherings in particular days. The results are considered signi�cant (P<0.05). *indicatesp<0.05 vs normal. &indicates p<0.05 vs VPA. #indicates p<0.05 vs VPA.
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Figure 8
TBR of normal control, VPA, VPA+O.S (200mg/kg), VPA+O.S (300mg/kg). The values are mean ± SEM(n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferroni posthoc test for all gatherings in particular days. The results are considered signi�cant (P<0.05). *indicatesp<0.05 vs normal. &indicates p<0.05 vs VPA. #indicates p<0.05 vs VPA.
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Figure 9
DBR of normal control, VPA, VPA+O.S (200mg/kg), VPA+O.S (300mg/kg). The values are mean ± SEM(n=6). Statistical analysis was done one-way analysis of variance (ANOVA) followed by Bonferroni’s posthoc test for all gatherings in particular days. The results are considered signi�cant (P<0.05). *indicatesp<0.05 vs normal. &indicates p<0.05 vs VPA. #indicates p<0.05 vs VPA.
